Trypanosoma brucei is a single cell protozoan parasite that infects humans and cattle in parts of Africa. Humans infected with T. brucei suffer from a severe neurological disease called African sleeping sickness that is inadvertently fatal when left untreated. Chemotherapeutic drugs against this parasite are few, relatively toxic, and logistically difficult to administer. The need to identify new drug targets and develop better medications is therefore urgent. This proposal will identify unique aspects of gene regulation in T. brucei, discovering novel mechanisms as potential drug targets. Unlike other eukaryotes, genes transcribed by RNA polymerase II (Pol II) in trypanosomes are arranged in large polycistronic transcription units (PTU) as long primary RNAs. Little is known about regulation of transcription initiation or termination. However, recent evidence indicates that epigenetic factors and especially base J, a hyper-modified nucleotide plays a significant role in regulation of Pol II mediated transcription. We have identified regulation of transcription initiation as well as evidence of regulated transcription termination. I propose two aims to study these modes of repression and the epigenetic mechanisms that are involved.